Basic Information

Common Name
B-hPD-L1 plus/hCEA MC38
Aliases
PD-L1: CD274, PDCD1L1;   CEA: CD66e, CEACAM5
Catalog Number
320722
Cell Type/ Disease
Colon Carcinoma
Species/ Strain
Mus musculus, C57BL/6
Notice
Biocytogen provides engineering service to generate the humanized tumor cell lines from customer’s own cell lines. Our humanized tumor cell lines are for pharmacology services ONLY.

Description

The mouse Pdl1 gene and Ceacam5 gene were replaced by human PD-L1 and CEACAM5 coding sequence in B-hPD-L1 plus/hCEA MC38 cells. Human PD-L1 and CEACAM5 are highly expressed on the surface of B-hPD-L1 plus/hCEA MC38 cells.

Application

B-hPD-L1 plus/hCEA MC38 cells have the capability to establish tumors in vivo and can be used for efficacy studies.

Target Strategy

  • The exogenous promoter and human PD-L1 coding sequence was inserted to replace part of murine exon 3. The insertion disrupts the endogenous murine Pdl1 gene, resulting in a non-functional transcript.
  • The exogenous promoter and human CEACAM5 coding sequence was inserted into the ROSA26 locus in murine.

Protein expression analysis

PD-L1 and CEACAM5 expression analysis in B-hPD-L1 plus/hCEA MC38 cells by flow cytometry. Single cell suspensions from B-hPD-L1 MC38 plus and B-hPD-L1 plus/hCEA MC38 cultures were stained with species-specific anti-PD-L1 and anti-CEACAM5 antibodies. Human PD-L1 was detected on the surface of B-hPD-L1 plus/hCEA MC38 and B-hPD-L1 MC38 plus cells. Human CEACAM5 was detected on the surface of B-hPD-L1 plus/hCEA MC38 cells. The 2-G09 clone of B-hPD-L1 plus/hCEA MC38 cells was used for in vivo experiments.

Tumor growth curve

Subcutaneous homograft tumor growth of B-hPD-L1 plus/hCEA MC38 cells. B-hPD-L1 plus/hCEA MC38 cells (5×105)  and B-hPD-L1 MC38 plus cells (5×105) were subcutaneously implanted into C57BL/6 mice (female, 5-8-week-old, n=5). Tumor volume and body weight were measured twice a week. (A) Average tumor volume ± SEM. (B)  Body weight (Mean± SEM). Volume was expressed in mm3 using the formula: V=0.5 X long diameter X short diameter2. As shown in panel A, B-hPD-L1 plus/hCEA MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.